Compositions and methods for inhibiting expression of transthyretin
View Patent ↗The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a transthyretin (TTR) gene, and methods of using the dsRNA to inhibit expression of TTR.
1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of transthyretin (TTR), wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region complementary to a part of a mRNA encoding transthyretin (TTR), wherein said region of complementarity is 19 nucleotides in length in length and the antisense strand comprises SEQ ID NO:170 and each strand of the dsRNA is 19, 20, 21, 22, 23, or 24 nucleotides in length.
2. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:449 and the antisense strand consists of SEQ ID NO:450.
3. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:729 and the antisense strand consists of SEQ ID NO:730.
4. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:1009 and the antisense strand consists of SEQ ID NO:1010.
5. The dsRNA of claim 1 , wherein the region of complementary consists of SEQ ID NO:169.
6. The dsRNA of claim 1 , wherein each strand is 21 nucleotides in length.
7. The dsRNA of claim 1 , wherein the dsRNA does not cleave a TTR mRNA between the adenine nucleotide at position 637 of SEQ ID NO:1331 and the guanine nucleotide at position 638 of SEQ ID NO:1331.
8. The dsRNA of claim 1 , wherein the dsRNA cleaves a TTR mRNA between the guanine nucleotide at position 636 of SEQ ID NO:1331 and the adenine nucleotide at position 637 of SEQ ID NO:1331.
9. The dsRNA of claim 1 , wherein the dsRNA anneals to a TTR mRNA between the guanine nucleotide at position 628 of SEQ ID NO:1331 and the uracil nucleotide at position 646 of SEQ ID NO: 1331.
10. The dsRNA of claim 1 , wherein said dsRNA comprises at least one modified nucleotide.
11. The dsRNA of claim 10 , wherein at least one of said modified nucleotides is chosen from the group of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.
12. The dsRNA of claim 10 , wherein said modified nucleotide is chosen from the group of: a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.
13. The dsRNA of claim 10 , comprising at least one 2′-O-methyl modified nucleotide.
14. The dsRNA of claim 1 , wherein the dsRNA is conjugated to a ligand.
15. The dsRNA of claim 1 , wherein administration of the dsRNA to a cell results in about 95% inhibition of TTR mRNA expression as measured by a real time PCR assay, wherein the cell is a HepG2 cell or a Hep3B cell, and wherein the concentration of the dsRNA is 10 nM.
16. The dsRNA of claim 1 , wherein said antisense strand base pairs with the guanine at position 628 of SEQ ID NO:1331.
17. A cell containing the dsRNA of claim 1 .
18. A vector comprising a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .
19. A cell comprising the vector of claim 18 .
20. A pharmaceutical composition for inhibiting expression of a TTR gene comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
21. A method of inhibiting TTR expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TTR gene, thereby inhibiting expression of the TTR gene in the cell.
22. A pharmaceutical composition for inhibiting expression of a TTR gene comprising the dsRNA of claim 4 and a pharmaceutically acceptable carrier.
23. A method of inhibiting TTR expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA of claim 4 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TTR gene, thereby inhibiting expression of the TTR gene in the cell.